title: Quantitation of glutathione by liquid chromatography/positive electrospray ionization tandem mass spectrometry.
abstract: Glutathione (GSH) is a tripeptide composed of glutamate, cysteine, and glycine. It is present in practically all cells and has several important roles, such as preventing the oxidation of the sulfhydryl groups of proteins within a cell. Evidence for GSH deficiency or depletion has been found in a variety of diseases and toxicity-related studies, including diabetes and induction of oxidative stress to form reactive oxygen species which cause DNA, lipid, and protein oxidations. A simple, selective, and sensitive analytical method for measuring low levels of GSH in biological fluids would therefore be desirable to conduct GSH deficiency or depletion-related mechanistic toxicity studies. Here a method for both low- and high-level quantitation of GSH from cultured cells and rat liver tissues via liquid chromatography/positive electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) has been developed. The lower limit of quantitation (LOQ) of the method was 5 ng/mL. The method is linear over a wide dynamic concentration range of 5.0 to 5000.0 ng/mL, with a correlation coefficient R2 > 0.99. The intra-day assay precision relative standard deviation (RSD) values for all quality control (QC) samples were < or =16.31%, with accuracy values ranging from 94.13 to 97.80%. The inter-day assay precision RSD values for all QC samples were < or =15.94%, with accuracy values ranging from 94.51 to 100.29%. With this method, low levels of GSH from diethyl maleate (DEM)-treated mouse lymphoma cells, and GSH in rat liver tissues, were quantified.
authors.affiliation: Toxicology and Environmental Research & Consulting, The Dow Chemical Company, 1803 Building, Midland, MI 48674, USA. fzzhang@dow.com
authors.forename: Fagen
authors.initials: F
authors.lastname: Zhang
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Bartels
authors.forename: David R
authors.initials: DR
authors.lastname: Geter
authors.forename: Yo-Chan
authors.initials: YC
authors.lastname: Jeong
authors.forename: Melissa R
authors.initials: MR
authors.lastname: Schisler
authors.forename: Amanda J
authors.initials: AJ
authors.lastname: Wood
authors.forename: Lynn
authors.initials: L
authors.lastname: Kan
authors.forename: B Bhaskar
authors.initials: BB
authors.lastname: Gollapudi
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002853:Chromatography, Liquid / Q000379:methods*; D005978:Glutathione / Q000032:analysis*; D008099:Liver / Q000737:chemistry; D008297:Male; D051379:Mice; D051381:Rats; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D005978:Glutathione